Quantification and sizing of exosomes using NanoSight NS300 particle counter and analysis of protein exosome markers by western blotting.

For “Fig 3”, panels A, B and C, x-axis represents particle size and y-axis represents the mode peak particle concentration. Mode peak concentration value is less than the total particle concentration. The total particle concentration is represented by the areas under the curves. Software used in the instrument automatically calculate the areas under the curves and gives estimated total particle concentration. Since sample was diluted 5-times, concentration value provided by software was multiplied by five. A, Exosomes were analyzed under light scatter mode. Size is heterogeneous ranging from 30–260 nm. Mean size is 92.6 nm and mode is 39.7 nm. Concentration 9.25 × 10⁹ particles/mL. B, Analysis of Exo-FTIC™ (green fluorescence) labeled exosomes using fluorescence mode. Exosomes are heterogeneous in size ranging from 30–260 nm. Mean size is 113.3 nm and mode is 51.6 nm. Concentration 24 × 10⁹ particles/mL plasma. C, Analysis of exosomes stained with Quant-iT™ PicoGreen^® dsDNA reagent using fluorescence mode. Exosomes are heterogeneous in size ranging from 30–260 nm. Mean size is 106.5 nm and mode is 45.5 nm. Concentration 18 × 10⁹ particles/mL plasma.